Glycolysis

 Greek word- glycos meaning sweet/sugar and lysis meaning dissolution

 Glycolysis is the first step in the breakdown of glucose to extract energy

for cellular metabolism. Glycolysis is a series of reactions that extract
energy from glucose by splitting it into two three-carbon molecules called
pyruvates.

 Sequence of reactions converting glucose to pyruvate with production of

ATP.

 Embden-Meyerhof pathway
 In organisms that perform cellular respiration, glycolysis is the first stage of
this process. However, glycolysis doesn’t require oxygen, and many
anaerobic organisms—organisms that do not use oxygen—also have this
pathway.

 Glycolysis takes place in the cytosol of a cell, and it can be broken down into
two main phases:
 energy-requiring phase,
 energy-releasing phase,

 Reversal of glycolysis along with the alternate arrangement at the

irreversible steps will result in the synthesis of glucose (Gluconeogenesis)
 Energy-requiring phase- In this phase, the starting
molecule of glucose gets rearranged, and two phosphate
groups are attached to it. The phosphate groups make the
modified sugar—now called fructose-1,6-bisphosphate—
unstable, allowing it to split in half and form two
phosphate-bearing three-carbon sugars. Because the
phosphates used in these steps come from ATP, 2
ATP molecules get used up.
 Splitting phase- In this step,
fructose – 1,6-diphosphate get
dissociated to get glyceraldehyde-
3-phosphate and dihydroxyacetone
phosphate
 Energy-releasing phase - In this phase, each
three-carbon sugar is converted into another
three-carbon molecule, pyruvate, through a
series of reactions. In these reactions, two
ATP molecules and one  NADH molecule are
made. Because this phase takes place twice,
once for each of the two three-carbon sugars,
it makes four ATP and two NADH overall.
 Overall, glycolysis converts one six-carbon
molecule of glucose into two three-carbon
molecules of pyruvate. The net products of
this process are two molecules of ATP and two
molecules of NADH.
The overall reaction of glycolysis which occurs in the
cytoplasm is represented simply as:

C6H12O6 + 2 NAD+ + 2 ADP + 2 P —–> 2 pyruvic

acid, (CH3(C=O)COOH + 2 ATP + 2 NADH + 2 H+
Step 1. Hexokinase

The first step in glycolysis is the conversion of D-glucose into glucose-6-phosphate.

The enzyme that catalyzes this reaction is hexokinase. 

Glucose

ATP

Hexokinase &
Mg2+ ADP

Glucose-6- phosphate
Pi
Step 2. Phosphoglucose Isomerase

The second reaction of glycolysis is the rearrangement of glucose 6-phosphate (G6P) into
fructose 6-phosphate (F6P) by glucose phosphate isomerase (Phosphoglucose Isomerase).

Glucose-6- phosphate
Pi

Phosphoglucose
Isomerase

Fructose-6-
Pi phosphate
Step 3. Phosphofructokinase

Phosphofructokinase, with magnesium as a cofactor, changes fructose 6-phosphate into fructose 1,6-
bisphosphate.

Fructose-6-
Pi
phosphate

ATP

Phosphofructokinase & Mg2+

ADP

Fructose-1,6-
Pi Pi
phosphate
Step 4. Aldolase

The enzyme Aldolase splits fructose 1, 6-bisphosphate into two sugars that are isomers of each
other. These two sugars are dihydroxyacetone phosphate  (DHAP) and glyceraldehyde 3-
phosphate (GAP).

Fructose-1,6-
Pi Pi
phosphate

Fructose bisphosphate aldolase

Pi isomer Pi
ase

DHAP GAP
Step 5. Triosephosphate isomerase

The enzyme triosephosphate isomerase rapidly inter- converts the molecules dihydroxyacetone phosphate
(DHAP) and glyceraldehyde 3-phosphate (GAP). Glyceraldehyde phosphate is removed / used in next step of
Glycolysis.

Pi

DHAP

Triosephosphate isomerase

Pi

GAP
Step 6. Glyceraldehyde-3-phosphate Dehydrogenase

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) dehydrogenates and adds an inorganic phosphate to

glyceraldehyde 3-phosphate, producing 1,3-bisphosphoglycerate.

Pi

GAP

Pi NAD+
2e

Triosephosphate isomerase NADH H+

Pi Pi
Step 7. Phosphoglycerate kinase

Phosphoglycerate kinase transfers a phosphate group from 1,3-bisphosphoglycerate to ADP to form ATP and
3-phosphoglycerate.

Pi Pi

ADP

Phosphoglycerate kinase
ATP

Pi
Step 8. Phosphoglycerate Mutase

The enzyme phosphoglycero mutase relocates the P from 3- phosphoglycerate from the 3rd carbon to the 2nd
carbon to form 2-phosphoglycerate.

Pi

Phosphoglycerate
mutase

Pi
Step 9. Enolase

The enzyme enolase removes a molecule of water from 2-phosphoglycerate to form phosphoenolpyruvic
acid (PEP).

Pi

Phosphoglycerate H2O
mutase

Pi
Step 10. Pyruvate kinase

PEP readily donates its phosphate group to ADP , making a second molecule of ATP. As it loses its
phosphate, PEP is converted to pyruvate, the end product of glycolysis.

Pi

ADP

Pyruvate kinase
ATP